Method of screening for colon cancer using biomarkers

Abstract

The present invention relates to biomarkers for colon cancers, specifically adenomas and adenocarcinomas in the GI tract. The inventors have discovered that the expression and/or overexpression of biomarkers such as CLDN1, GPR56, GRM8, LY6G6D, TLR4 and SLCO1B3 are indicative of adenomas and adenocarcinomas. A method of detecting colon cancer using targeted molecular imaging agents is also presented.

What is claimed is: 
     
       1. A method of detecting colon cancer in a patient comprising:
 obtaining a gastrointestinal tract sample suspected of containing a neoplasm from a patient; 
 detecting the presence of at least one biomarker in the sample by contacting the sample with an antibody selected from Table 2 wherein the biomarker is selected from the group consisting of claudin 1 (CLDN1), G protein-coupled receptor (GPR56), glutamate receptor, metabotropic 8 (GRM8), lymphocyte antigen 6 complex, locus G6D (LY6G6D), toll-like receptor 4 (TLR4) and solute carrier organic anion transporter family member 1B3 (SLCO1B3); 
 detecting a complex between the antibody and the at least one biomarker; 
 generating a pathology score for the at least one biomarker wherein the pathology score is based on staining intensity and degree of epithelial cell positivity; and 
 associating the pathology score with presence of colon cancer using statistical methods wherein the pathology score greater than or equal to 4 is indicative of colon cancer. 
 
     
     
       2. The method of  claim 1 , wherein the at least one biomarker is a combination of CLDN1, LY6G6D and TLR4. 
     
     
       3. The method of  claim 1 , wherein the at least one biomarker is a combination of TLR4, GPR56 and CLDN1. 
     
     
       4. The method of  claim 1 , wherein the at least one biomarker is a combination of TLR4 and GRM8. 
     
     
       5. The method of  claim 1 , wherein the at least one biomarker is CLDN1. 
     
     
       6. The method of  claim 1 , wherein the at least one biomarker is TLR4. 
     
     
       7. A method of detecting colon cancer comprising:
 obtaining an expression level of at least one biomarker in a sample by contacting the sample with an antibody selected from Table 2 wherein the biomarker is selected from the group consisting of claudin 1 (CLDN1), G protein-coupled receptor (GPR56), glutamate receptor, metabotropic 8 (GRM8), lymphocyte antigen 6 complex, locus G6D (LY6G6D), toll-like receptor 4 (TLR4) and solute carrier organic anion transporter family member 1B3 (SLCO1B3) in a patient sample; 
 detecting a complex between the antibody and the at least one biomarker; 
 generating a pathology score for the at least one biomarker based on staining intensity and degree of epithelial cell positivity; and 
 comparing the pathology score of the at least one biomarker to a predetermined control pathology score; 
 wherein a higher pathology score of the at least one biomarker as compared to the predetermined control pathology score indicates the presence of colon cancer. 
 
     
     
       8. The method of  claim 7 , wherein the at least one biomarker is a combination of CLDN1, LY6G6D and TLR4. 
     
     
       9. The method of  claim 7 , wherein the at least one biomarker is a combination of TLR4, GPR56 and CLDN1. 
     
     
       10. The method of  claim 7 , wherein the at least one biomarker is a combination of TLR4 and GRM8. 
     
     
       11. The method of  claim 7 , wherein the at least one biomarker is CLDN1. 
     
     
       12. The method of  claim 7 , wherein the at least one biomarker is TLR4. 
     
     
       13. A method of screening for colon cancer comprising:
 providing at least one magnetic resonance imaging (MRI) contrast agent wherein the contrast agent is a gadoteric acid (Gd)-sucrose construct; 
 binding the at least one contrast agent to at least one colorectal biomarker-specific synthetic ligand wherein the biomarker is selected from the group consisting of claudin 1 (CLDN1), G protein-coupled receptor (GPR56), glutamate receptor, metabotropic 8 (GRM8), lymphocyte antigen 6 complex, locus G6D (LY6G6D), toll-like receptor 4 (TLR4) and solute carrier organic anion transporter family member 1B3 (SLCO1B3); 
 adding the contrast agent with bound ligand to a pharmaceutically acceptable carrier to form a composition; 
 administering the composition to a patient; and 
 imaging the patient using radiation at least about 8.5 hours after administration; 
 wherein detection of the contrast agent with bound ligand in the patient is indicative of colon cancer. 
 
     
     
       14. The method of  claim 13 , wherein multivalent binding is used to bind multiple copies of the biomarker-specific ligand to the contrast agent. 
     
     
       15. The method of  claim 13 , wherein the at least one biomarker is a combination of CLDN1, LY6G6D and TLR4. 
     
     
       16. The method of  claim 13 , wherein the at least one biomarker is a combination of TLR4, GPR56 and CLDN1. 
     
     
       17. The method of  claim 13 , wherein the at least one biomarker is a combination of TLR4 and GRM8. 
     
     
       18. The method of  claim 13 , wherein the at least one biomarker is CLDN1. 
     
     
       19. The method of  claim 13 , wherein the at least one biomarker is TLR4. 
     
     
       20. The method of  claim 13 , wherein the Gd-sucrose construct is comprised of a sucrose scaffold conjugated with a plurality of Gd(III)-DOTA chelates. 
     
     
       21. The method of  claim 20 , wherein the number of chelates is between about 6 and about 8. 
     
     
       22. A method of screening for colon cancer comprising:
 providing at least one acid-stable and protease-resistant computed tomography (CT) contrast agent having a built-in attachment site for a targeting group wherein the contrast agent has a high binding affinity for and is selectively targeted to at least one colorectal cancer lesion; 
 developing at least one colorectal biomarker-specific synthetic ligand wherein the biomarker is selected from the group consisting of claudin 1 (CLDN1), G protein-coupled receptor (GPR56), glutamate receptor, metabotropic 8 (GRM8), lymphocyte antigen 6 complex, locus G6D (LY6G6D), toll-like receptor 4 (TLR4) and solute carrier organic anion transporter family member 1B3 (SLCO1B3); 
 binding the at least one contrast agent to the at least one colorectal biomarker-specific synthetic ligand wherein multivalent binding is used to bind multiple copies of the biomarker-specific ligand to the contrast agent; 
 adding the contrast agent with bound ligand to a pharmaceutically acceptable carrier to form a composition wherein the contrast agent and the bound ligand are stable in the digestive tract; 
 administering the composition to a patient; and 
 imaging the patient using radiation at least about 8.5 hours after administration; 
 wherein detection of the contrast agent with bound ligand in the patient is indicative of colon cancer; 
 wherein the at least one colorectal biomarker-specific synthetic ligand has a peptidomimetic scaffold. 
 
     
     
       23. The method of  claim 22 , wherein the composition is orally administered to the patient. 
     
     
       24. The method of  claim 22 , wherein the at least one contrast agent is a plurality of gold nanoparticles between about 1.5 nm to about 6 nm in size. 
     
     
       25. The method of  claim 22 , wherein the at least one biomarker is a combination of CLDN1, LY6G6D and TLR4. 
     
     
       26. The method of  claim 22 , wherein the at least one biomarker is a combination of TLR4, GPR56 and CLDN1. 
     
     
       27. The method of  claim 22 , wherein the at least one biomarker is a combination of TLR4 and GRM8.